Production of cell cultured meat based on the optimization of the co-differentiation conditions of adipose precursor cells and muscle stem cells

• Combining food grade soybean oil and Asc-2P derivatives effectively enhances lipid droplet formation, accelerating APCs differentiation. • Reducing serum in the differentiation medium of APCs and co-differentiating MuSCs with APCs at a 7:3 ratio can simultaneously produce a large number of muscle tubes and lipid droplets. • 3D-printed CCM based on the co-differentiation of APCs and MuSCs has a similar appearance to traditional meat and similar flavor profiles in volatile flavor substances. Cell cultured meat (CCM) has become a research hotspot in the field of meat substitutes because of its environmental friendliness. Improving the overall flavor of CCM and further enhancing consumer acceptance is the key to its entry into the market for sale. Therefore, this study aimed to develop differential medium with higher food safety for adipose precursor cells (APCs) and optimize the co-differentiation conditions of APCs and muscle stem cells (MuSCs) on this basis. Then, using co-differentiated APCs and MuSCs as 3D printed bioinks to produce CCM. The results of this study indicate that food grade soybean oil and l-Ascorbic acid 2-phosphate (Asc-2P) can significantly promote lipid droplet generation in APCs and shorten APCs differentiation time ( P <0.05). Simultaneous differentiation of MuSCs and APCs was achieved using a 7:3 inoculation ratio combined with optimization of the co-differentiation medium. And the 3D-printed CCM has a similar appearance to traditional meat and similar flavor profiles in volatile flavor substances such as alkanes, alcohols and aliphatic compounds. Ultimately, this study provides a theoretical support for the production of higher safety and better flavored CCM.